Muhammad Yasir

Culture of previously uncultured members of the human gut microbiota by culturomics

Metagenomics revolutionized the understanding of the relations among the human microbiome, health and diseases, but generated a countless number of sequences that have not been assigned to a known microorganism1. The pure culture of prokaryotes, neglected in recent decades, remains essential to elucidating the role of these organisms2. We recently introduced microbial culturomics, a culturing approach that uses multiple culture conditions and matrix-assisted laser desorption/ionization–time of flight and 16S rRNA for identification2. Here, we have selected the best culture conditions to increase the number of studied samples and have applied new protocols (fresh-sample inoculation; detection of microcolonies and specific cultures of Proteobacteria and microaerophilic and halophilic prokaryotes) to address the weaknesses of the previous studies3–5. We identified 1,057 prokaryotic species, thereby adding 531 species to the human gut repertoire: 146 bacteria known in humans but not in the gut, 187 bacteria and 1 archaea not previously isolated in humans, and 197 potentially new species. Genome sequencing was performed on the new species. By comparing the results of the metagenomic and culturomic analyses, we show that the use of culturomics allows the culture of organisms corresponding to sequences previously not assigned. Altogether, culturomics doubles the number of species isolated at least once from the human gut.

Bacterial community composition and chitinase gene diversity of vermicompost with antifungal activity

Bacterial communities and chitinase gene diversity of vermicompost (VC) were investigated to clarify the influence of earthworms on the inhibition of plant pathogenic fungi in VC. The spore germination of Fusarium moniliforme was reduced in VC aqueous extracts prepared from paper sludge and dairy sludge (fresh sludge, FS). The bacterial communities were examined by culture-dependent and -independent analyses. Unique clones selected from 16S rRNA libraries of FS and VC on the basis of restriction fragment length polymorphism (RFLP) fell into the major lineages of the domain bacteria Proteobacteria, Bacteroidetes, Verrucomicrobia, Actinobacteria and Firmicutes. Among culture isolates, Actinobacteria dominated in VC, while almost equal numbers of Actinobacteria and Proteobacteria were present in FS. Analysis of chitinolytic isolates and chitinase gene diversity revealed that chitinolytic bacterial communities were enriched in VC. Populations of bacteria that inhibited plant fungal pathogens were higher in VC than in FS and particularly chitinolytic isolates were most active against the target fungi.

Role of gut microbiota in obesity, type 2 diabetes and Alzheimer's disease.

In recent years, there is a growing interest in research to investigate the importance of gut microbiome in health and diseases. This opens a new area of research for the role of microbial flora of the human gut in inflammation, energy homeostasis, pathogenesis of obesity and other associated disorders. Recent studies propose association of the gut microbiome with development of obesity and metabolic syndromes, such as type 2 diabetes mellitus (T2DM). The T2DM is a metabolic disease that is mainly caused by obesity-linked insulin resistance. The vascular effects of obesity appears to play a role in the development of Alzheimers disease (AD) that is one of the rapidly growing diseases of a late stage of life all over the world. Studies from both humans and mice models have been demonstrated the engagement of gut microbial flora in the pathogenesis of obesity and host metabolism. The aim of this review is to discuss the current findings that may explain the cascade of gut microbial flora participation in the development of obesity, T2DM and further initiation of AD. In addition, the available data regarding the mechanisms that have been proposed to elucidate the role of gut microbiota in weight gain and possible cause of T2DM and AD have been examined.

Diversity and Characterization of Endophytic Bacteria Associated with Tidal Flat Plants and their Antagonistic Effects on Oomycetous Plant Pathogens

Endophytic bacterial communities of tidal flat plants antagonistic to oomycete plant pathogens were studied by the isolation of 256 root colonizing endophytic bacteria from surface-disinfected root tissues of six plants (Rosa rugosa, Suaeda maritima, Vitex rotundifolia, Carex scabrifolia, Glehnia littoralis and Elymus mollis) growing in a tidal flat area of Namhae Island, Korea. To understand the antagonistic potential, an in vitro antagonistic assay was performed to characterize and identify strains that were antagonistic to the oomycete plant pathogens Phytophthora capsici and Pythium ultimum from the total population. Nine percent of the total number of isolated bacteria exhibited in vitro inhibitory activity against target plant pathogenic oomycetes. Taxonomic and phylogenetic placement of the antagonistic bacteria was investigated by analysis of the 16S rRNA gene sequences. The sequence analysis classified the antagonistic strains into four major classes of the domain bacteria (Firmicutes, α-Proteobacteria, γ-Proteobacteria and Actinomycetes) and 10 different genera. Further production of secondary metabolites, hydrolytic enzymes and plant growth promoting traits were determined for the putative new species of antagonistic endophytic bacteria. These new strains could not be identified as known species of α-Proteobacteria, and so may represent novel bacterial taxa. The unexpected high antagonistic bacterial diversity associated with the tidal flat plants may be indicative of their importance in tidal flat plants as a promising source of novel antimicrobial compounds and biocontrol agents.

Lysobacter oryzae sp. nov., isolated from the rhizosphere of rice (Oryza sativa L.)

The taxonomic position of a novel bacterial strain, YC6269(T), isolated from the rhizosphere of rice (Oryza sativa L.) managed under no-tillage practice in Jinju, South Korea, was studied using polyphasic approach. Cells of the strain were Gram-negative, rod-shaped and facultatively anaerobic. The novel strain grew at a temperature of 15-42 degrees C (optimum at 28 degrees C). Growth of the strain occurred between pH 5.5 and 11.0, with an optimum at pH 7.0-8.0. The G+C content of the total DNA was 67.4 mol%. The 16S rRNA gene sequence of the strain was most closely related to species of the genus Lysobacter, Lysobacter yangpyeongensis DSM 17635(T) (98.6 %), Lysobacter niabensis GH34-4(T) (97.2 %), Lysobacter enzymogenes DSM 2043(T) (96.9 %), Lysobacter daejeonensis DSM 17634(T) (96.3 %) and Lysobacter niastensis GH41-7(T) (96.2 %). The novel strain showed <96.0 % similarity with other species of the genus Lysobacter. Chemotaxonomic data (major quinone, Q-8; major polar lipids, phosphatidylethanolamine, phosphatidylglycerol and phosphatidyl-N-methylethanolamine, and major fatty acids, C(15 : 0) iso, C(16 : 0) iso, C(17 : 0) iso and C(17 : 1) iso omega9c) supported the affiliation of strain YC6269(T) to the genus Lysobacter. Phylogenetic analysis based on the 16S rRNA gene sequences, DNA-DNA hybridization data and biochemical and physiological characteristics strongly supported the genotypic and phenotypic differentiation of strain YC6269(T) from recognized species of the genus Lysobacter. Strain YC6269(T), therefore, represents a novel member of the genus Lysobacter, for which the name Lysobacter oryzae sp. nov. is proposed. The type strain is YC6269(T) (=KCTC 22249(T)=DSM 21044(T)).

Comparison of the gut microbiota of people in France and Saudi Arabia

Background/Objectives:The gut microbiota contributes to energy acquisition from food, and changes in the gut microbiome are associated with obesity. The eating habits of Saudis are much different than those of Europeans, and our objective was to compare the fecal microbiota of obese and normal weight Saudis and French.Subjects/Methods:Illumina MiSeq deep sequencing was used to test the gut microbiota of 9 normal weight and 9 obese individuals from Saudi Arabia and 16 normal weight and 12 obese individuals from France.Results:Obese French possessed significantly more relative Proteobacteria (P=0.002) and Bacteroidetes (P=0.05) and had lower richness and biodiversity at all the operational taxonomic unit (OTU) cutoffs (P<0.05) than normal weight French. Obese Saudis possessed significantly more Firmicutes (P=0.001) without a difference in richness (P=0.2) and biodiversity (P=0.3) compared with normal weight Saudis. We found a common bacterial species core of 23 species existing in ⩾50% of obese and normal weight Saudis and 29 species in ⩾50% of obese and normal weight French. Actinomyces odontolyticus, Escherichia coli and Ruminococcus obeum were present in at least 50% of all individuals tested. French individuals had significantly higher richness and biodiversity compared with Saudis at all the OTU cutoffs (P<0.05).Conclusion:Microbiota differences between obese and normal weight French were not similar to those between obese and normal weight Saudis. The studies of different populations can result in contrasting data regarding the associations of the gut microbiota and obesity.

Nocardioides caricicola sp. nov., an endophytic bacterium isolated from a halophyte, Carex scabrifolia Steud.

A Gram-staining-positive, coccoid to rod-shaped bacterium, designated strain YC6903(T), was isolated from a halophytic plant (Carex scabrifolia Steud.) collected from sand dunes at Namhae Island, Korea, and its taxonomic position was investigated by using a polyphasic approach. Strain YC6903(T) grew optimally at 30 °C and at pH 8.0. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain YC6903(T) belongs to the genus Nocardioides in the family Nocardioidaceae. Strain YC6903(T) was related most closely to Nocardioides pyridinolyticus OS4(T) (97.0u200a% 16S rRNA gene sequence similarity), Nocardioides dokdonensis FR1436(T) (96.6u200a%), Nocardioides aquiterrae GW-9(T) (96.6u200a%) and Nocardioides hankookensis DS-30(T) (96.6u200a%). The cell-wall peptidoglycan contained LL-diaminopimelic acid and MK-8(H(4)) was the major respiratory quinone. The mean (±SD) level of DNA-DNA relatedness between strain YC6903(T) and N. pyridinolyticus OS4(T) was 53.5±5.5u200a%. The predominant cellular fatty acid of strain YC6903(T) was iso-C(16u200a:u200a0) (28.9u200a%). The DNA G+C content was 71.7 mol%. Phenotypic, phylogenetic and chemotaxonomic data indicated that strain YC6903(T) represents a novel species of the genus Nocardioides, for which the name Nocardioides caricicola sp. nov. is proposed. The type strain is YC6903(T) (=KACC 13778(T) =DSM 22177(T)).

Link Between Chronic Bacterial Inflammation and Alzheimer Disease

Alzheimers disease (AD) is a degenerative disease of brain that is associated with dementia, brain atrophy, accumulation of hyperphosphorylated tau protein and amyloid-beta peptide in hippocampus and cortex region of the brain. The development of AD is a multifactorial process that may also involve infection with bacterial pathogens. Recent studies suggest that bacteria including spirochetes have the potential to initiate cascade of events, leading to inflammatory condition of the central nervous system. Bacteria and spirochetes are activators of proinflammatory cytokines, generate free radicals, nitric oxide and further induction of apoptosis. Infection with these microbes may be considered as a risk factor for pathophysiology of AD or to cognitive changes. Recent studies have revealed that exposure to these microorganisms induces Aβ accumulation and tau protein phosphorylation, and chronic infections with these pathogenic bacteria can possibly contribute to progression of AD. In this article, we update and review the role of bacteria in the pathogenesis of AD resulting from initiation of cascade events in chronic inflammations and amyloidogenesis. Controlling these chronic infections with antibacterial or anti-inflammatory drugs will allow preventing inflammation, a risk factor for AD.

Chitinophaga eiseniae sp. nov., isolated from vermicompost.

A Gram-negative, rod-shaped bacterial strain, YC6729(T), was isolated from vermicompost collected at Masan, Korea, and its taxonomic position was investigated by a polyphasic taxonomic approach. Strain YC6729(T) grew optimally at 30 °C and at pH 6.5-8.5. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain YC6729(T) belongs to the genus Chitinophaga in the family Chitinophagaceae. It was related most closely to Chitinophaga terrae KP01(T) (96.4u200a% 16S rRNA gene sequence similarity), Chitinophaga ginsengisegetis Gsoil 040(T) (96.1u200a%), Chitinophaga arvensicola IAM 12650(T) (96.1u200a%) and Chitinophaga pinensis DSM 2588(T) (93.3u200a%). Strain YC6729(T) contained MK-7 as the major menaquinone and homospermidine as the major polyamine. The fatty acids of strain YC6729(T) were iso-C(15u200a:u200a0), C(16u200a:u200a1)ω5c, iso-C(17u200a:u200a0) 3-OH, C(16u200a:u200a0), anteiso-C(18u200a:u200a0) and/or C(18u200a:u200a2)ω6,9c, iso-C(15u200a:u200a0) 2-OH and/or C(16u200a:u200a1)ω7c, C(14u200a:u200a0), iso-C(15u200a:u200a0) 3-OH, iso-C(15u200a:u200a1) G, C(18u200a:u200a1)ω5c, iso-C(15u200a:u200a1) I and/or C(13u200a:u200a0) 3-OH, C(13u200a:u200a0) 2-OH, C(16u200a:u200a0) 3-OH and unknown fatty acid ECL 13.565. The polar lipid profile contained phosphatidylethanolamine, unknown aminolipids and unknown lipids. The total DNA G+C content of strain YC6729(T) was 48.9 mol%. The phenotypic, chemotaxonomic and phylogenetic data showed that strain YC6729(T) represents a novel species of the genus Chitinophaga, for which the name Chitinophaga eiseniae sp. nov. is proposed. The type strain is YC6729(T) (u200a=u200aKACC 13774(T) u200a=u200aDSM 22224(T)).

Paper money and coins as potential vectors of transmissible disease.

Paper currency and coins may be a public health risk when associated with the simultaneous handling of food and could lead to the spread of nosocomial infections. Banknotes recovered from hospitals may be highly contaminated by Staphylococcus aureus. Salmonella species, Escherichia coli and S. aureus are commonly isolated from banknotes from food outlets. Laboratory simulations revealed that methicillin-resistant S. aureus can easily survive on coins, whereas E. coli, Salmonella species and viruses, including human influenza virus, Norovirus, Rhinovirus, hepatitis A virus, and Rotavirus, can be transmitted through hand contact. Large-scale, 16S rRNA, metagenomic studies and culturomics have the capacity to dramatically expand the known diversity of bacteria and viruses on money and fomites. This review summarizes the latest research on the potential of paper currency and coins to serve as sources of pathogenic agents.